Stathmin is highly expressed in eutopic endometrium of patient with endometriosis

Objective: To investigate the expression of stathmin in eutopic endometrium of women with or without endometriosis.Methods: The eutopic endometrium samples were collected from thirty-six women with endometriosis and nineteen age-matched women without endometriosis. The expression of stathmin protein was detected by immunohisto-chemistry and Western blot.Results: Stathmin was expressed in both endometrial glandular and stromal cells throughout the menstrual cycle. Stathmin expression had no significant difference between proliferative and secretory phase. Stathmin was overexpressed in eutopic endometrium of women with endometriosis compared with those without endometriosis.Conclusion: The overexpressed stathmin in endometrium of patient with endometriosis may play a certain role in the pathogenesis of endometriosis.     

低氧环境下子宫内膜基质细胞增殖及Prohibitin表达

目的观察常氧和低氧环境下子宫内膜基质细胞(ESCs)的增殖,探讨抗增殖蛋白prohibitin(PHB)在子宫内膜异位症(EMs)病理机制中的作用。方法 10例正常内膜组织(正常对照组)和10例EMs患者在位内膜组织(在位内膜组)各分为两份,分别置于常氧(21%O_2)与低氧(1%O_2)环境中培养。运用BrdU方法检测细胞增殖,并以光密度值(OD)代表细胞的增殖能力;通过流式细胞技术检测细胞凋亡;细胞免疫荧光技术和蛋白质印记法(Western blot)检测细胞中PHB表达。结果在常氧环境,在位内膜组的OD值略高于正常对照组但无统计学差异(P0.05);而在低氧环境,在位内膜组的OD值[(1.400±0.659)]显著高于正常对照组[(0.590±0.021)](P0.001),且显著高于常氧环境的在位内膜OD值[(1.156±0.044)](P0.05)。无论是常氧还是低氧环境,在位内膜组的细胞凋亡率均显著低于对照组[分别为(4.117±0.450)%vs.(7.247±0.707)%及(4.983±0.393)%vs.(9.867±0.675)%],差异均有统计学意义(P0.05)。在常氧环境,在位内膜组ESCs的PHB表达[(0.747±0.026)]显著高于正常对照组[(0.590±0.021)](P0.05);在低氧环境,正常对照组和在位内膜组ESCs的PHB表达水平均较常氧环境的PHB表达水平降低[分别为(0.358±0.071)vs.(0.590±0.021)和(0.458±0.054)vs.(0.747±0.026)],且差异有统计学意义(P0.05)。结论正常子宫内膜组织表达PHB,EMs患者的在位内膜组织PHB表达升高;低氧环境下ESCs的PHB表达水平显著降低,可能与促进ESCs增殖、抑制ESCs凋亡相关,因此PHB在EMs病理机制中起重要作用。     

子宫内膜异位症患者雌、孕激素受体和Bcl-2、Bax表达的研究

目的探讨子宫内膜异位症雌孕激素受体、细胞凋亡相关基因的表达。方法用免疫组织化学方法及图像分析技术检测雌激素受体（ER）、孕激素受体（PR）、B细胞淋巴瘤／白血病基因-2（Bcl-2）、Bcl相关的X蛋白（Bax）在子宫内膜异位症患者异位及在位内膜细胞中的表达情况。结果异位内膜ER、PR、Bcl-2表达明显低于在位内膜（P〈0．05）,异位内膜ER、Bcl-2表达明显高于正常内膜（P〈0．05）,异位内膜Bax表达明显低于正常内膜（P〈0．05）,Bax在异位内膜与在位内膜表达差异无统计学意义（P〉0．05）。结论无论是增殖期还是分泌期,在位内膜ER、PR表达均高于异位内膜及正常内膜,在位内膜细胞持续低水平增殖。在位内膜及异位内膜中Bcl-2和Bax的表达均与子宫内膜周期性改变无关,不受卵巢激素调节。     

子宫内膜异位症子宫内膜及输卵管细胞对鼠胚胎发育的影响

目的探讨子宫内膜异位症(内异症)子宫内膜及输卵管上皮细胞对小鼠胚胎体外发育的影响。方法小鼠2细胞期胚胎202个与卵巢巧克力囊肿患者术中切除的子宫内膜及输卵管上皮细胞共培养(实验组),314个与非卵巢巧克力囊肿患者子宫内膜及输卵管上皮细胞共培养作为对照。结果体外培养72~96h后,与对照组输卵管上皮细胞及子宫内膜共培养时有25.7%和26.2%的2细胞期胚胎发育到扩张期囊胚,与实验组输卵管上皮细胞共培养有25.9%发育到扩张期囊胚,二者无显著差异(P>0.05),而子宫内膜仅为11.1%,二者有显著差异(P<0.05)。结论内异症患者的输卵管上皮细胞对鼠早期胚胎体外发育无影响;子宫内膜对胚胎发育有明显的抑制作用,这可能与内异症的生育率降低有关。     

子宫内膜异位症中上皮型钙粘蛋白表达及其意义

目的探讨上皮型钙粘蛋白（E-cad）与子宫内膜异位症发病的关系。方法采用双抗体夹心ABC—ELISA法检测30例子宫内膜异位症（内异症组）患者血清E—cad的表达量,免疫组化法检测异位内膜、在位内膜E-cad的表达量,分别与同期30例卵巢成熟性畸胎瘤患者（对照组）血清、子宫内膜进行比较。结果内异症组血清E—cad表达量显著高于对照组（P〈0．001）;内异症组在位及异位内膜E—cad表达均低于对照组内膜（P=0．004）,尤以异位内膜表达量更低。结论Bcad在内异症患者血清、在位和异位内膜中的异常表达,可能与内异症的发生、发展有关。     

Adenocarcinoma arising on colo-rectal endometriosis: clinico-pathological considerations

Colo-rectal endometriosis represents a rare pathological event. Exceptionally, a malignant colo-rectal endometrioid carcinoma arising from endometriosis has been reported. In the present case, we describe a 32 year-old woman with clinical and radiological signs of large bowel sub-occlusive syndrome. Histologically, a sigmoid endometrioid adenocarcinoma associated with submucosal foci of eutopic endometrium was observed. Moreover, endometriotic tissue was observed at the level of the colonic left flexura and appendix. Colo-rectal endometriosis must be considered among risk factors for the development of large bowel adenocarcinoma in young female patients.     

子宫内膜异位症患者在位内膜分泌RANTES及对单核细胞的趋化活性

目的了解子宫内膜异位症(Em)患者的在位内膜在分泌RANTES及对单核细胞的趋化活性是否有别于非Em患者子宫内膜。方法白细胞介素(IL)-1β刺激培养Em与非Em患者子宫内膜基质细胞,分别在刺激培养后12、24、36、48、60、72、84和96 h取上清酶联免疫吸附试验(ELISA)检测RANTES水平;Boyden小室趋化实验分析刺激培养上清液对单核细胞的趋化活性及RANTES趋化单核细胞的能力。结果Em在位内膜从刺激培养60 h始,RANTES分泌量及趋化单核细胞的能力显著高于非Em在位内膜;Em在位内膜基质细胞刺激培养60 h上清所趋化的单核细胞中有55%是由RAN-TES趋化而来。结论在相似的培养条件下,Em患者的在位内膜基质细胞RANTES的表达模式和趋化单核细胞的活性不同于非Em患者子宫内膜。     

人增生性瘢痕中整合素连接激酶表达及与血管生成的关系

目的 探讨不同生长期人类瘢痕中整合素连接激酶(ILK)表达及与血管生成的关系.方法 (1)将笔者单位2009年12月-2010年12月收治的15例烧伤瘢痕患者按瘢痕生长时间分为:小于6个月组、6～12个月组、大于12个月组,每组5例.采集各组瘢痕组织,用免疫组织化学法观察ILK的表达分布,实时荧光定量RT-PCR法检测ILK mRNA的表达水平.(2)取小于6个月组瘢痕组织,分离培养瘢痕微血管内皮细胞(MEC),免疫磁珠法纯化后用花青类荧光染料Cy3标记的凝血因子Ⅷ进行鉴定,以人皮肤Fb为对照.取对数生长期MEC,按随机数字表法分为3组:对照组,仅用含微血管生长添加剂的M131培养液培养;空质粒组,用空载质粒转染;ILK互补DNA转染组,用ILK互补DNA表达质粒转染.转染24h后,实时荧光定量RT-PCR法检测各组细胞中ILK、激酶功能区受体(KD R)、fms样酪氨酸激酶1(Flt1)的mRNA表达情况.对数据进行单因素方差分析. 结果 (1)小于6个月组瘢痕组织表皮基底细胞、MEC及Fb胞质中均有ILK阳性表达,6～12个月组瘢痕组织中仅表皮基底细胞可见ILK阳性表达,大于12个月组瘢痕组织中ILK阳性表达不明显.(2)小于6个月组瘢痕组织中ILK mRNA表达水平(0.34±0.16)明显高于6～12个月组(0.17±0.06)及大于12个月组(0.07±0.13),F=37.007,P - 0.000.(3)纯化后MEC呈铺路石样密集生长,胞质中凝血因子Ⅷ呈阳性表达;人皮肤Fb中未见凝血因子Ⅷ表达.(4)ILK互补DNA转染组瘢痕MEC中ILK、KDR及Flt-1的mRNA表达水平分别为57.807±5.556、0.836±0.014、0.162±0.005,均显著高于对照组的0.018±0.003、0.028±0.020、0.023±0.004和空质粒组的0.042±0.005、0.039±0.0070.046±0.003(F值分别为87.110、11.241、18.199,P值均小于0.01).结论 ILK主要表达于小于6个月的早期瘢痕组织中,并可以通过调节瘢痕MEC中的KDR及Flt-1 mRNA表达来影响早期瘢痕的血管生成,在早期瘢痕增生过程中具有重要作用.     

Accuracy of laparoscopic diagnosis of endometriosis.

BACKGROUND AND OBJECTIVE: Laparoscopy is the standard method to visually identify endometriotic lesions under magnification within and outside the minor pelvis. The aim of this study was to analyze the accuracy of laparoscopic visualization in diagnosing the various endometriotic sites as confirmed histologically. METHOD: Presumed endometriotic sites were observed in 164 patients operated on under the clinical suspicion of endometriosis. Targeted biopsies were performed for histologic corroboration, comparing the laparoscopic findings and diagnosis to the histological results. RESULT: The histological reports of the biopsies confirmed the presence of endometriosis in 138 patients (84.1%), but in 26 patients (15.9%), no evidence of endometriosis was observed. 100% of "red" lesions, 92% of "black" lesions, and 31% of "white" lesions turned out to be endometriosis. Of the 264 various suspected endometriotic sites observed, 142 (53.8%) were confirmed histologically. The most accurate diagnosis was in lesions on the parietal peritoneum of the pelvis, confirmed in 9/9 cases (100%); the ovarian fossa, confirmed in 8/12 cases (66.7%); and the uterosacral ligaments and posterior surface of the broad ligament, confirmed in 83/138 cases (60.1%). As for the other sites, the histologic confirmation rates in the ovarian surface, bowel serosa, and vesicouterine fold of the peritoneum were 48%, 40%, and 13%, respectively. CONCLUSION: Endometriosis has a multiple appearance, and the lesions may be confused with nonendometriotic lesions. It is clear that a nonhistology-based diagnosis may lead to unnecessary prolonged medical treatment and operations and may delay the proper treatment measures from being applied. Therefore, a meticulous histological confirmation should still be the first step in the laparoscopic diagnosis and treatment of suspected endometriosis.     

控制性卵巢刺激方案及血清CA125水平对复发性卵巢子宫内膜异位囊肿患者体外受精-胚胎移植结局的影响

目的研究卵巢刺激方案及血清CA125水平对手术后复发的卵巢子宫内膜异位囊肿不孕患者体外受精-胚胎移植（IVF—ET）治疗结果的影响。方法回顾分析2004年1月至2007年12月间经IVF治疗的39个周期（38例）卵巢子宫内膜异位囊肿手术后复发的不孕妇女治疗的临床结果,以及卵巢刺激方案与垂体降调后血清CA125水平间的相关性。结果长效促性腺激素释放激素激动剂（GnRH—a）垂体降调后采用短方案卵巢刺激13个周期,临床妊娠6个周期（46．2％）;长方案卵巢刺激26个周期,临床妊娠8个周期（30．8％）。长方案组周期平均获卵数（8．42±5．22）显著高于短方案组（5．00±2．35）（P=0．021）,但受精率、周期临床妊娠率组间无显著差异（P=0．482）。垂体降调后23个周期血清CA125为（18．5±12．6）U／ml;其中妊娠8例CA125为（10．2±5．8）U／ml,与妊娠结局呈显著负相关（r=0．492,P=0．017）。结论卵巢子宫内膜异位囊肿术后复发的不孕妇女,垂体降调后作IVF有较好疗效。卵巢刺激短方案或长方案的妊娠结局无显著差异。降调后血清CA125水平可作为疗效的预测指标。     

Expression of vascular endothelial growth factor and its receptors Flt-1 and KDR/Flk-1 in chronic cyclosporine nephrotoxicity

BACKGROUND: Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen involved in angiogenesis, wound healing, and inflammation. METHODS: Rats placed on low salt diet (LSD) or normal salt diet (NSD) were treated with cyclosporine (CsA) or vehicle (VH) and killed at 7 or 28 days. We studied the expression of VEGF and its receptors Flt-1 and KDR/Flk-1 mRNA by Northern and that of VEGF protein by Western blot. RESULTS: CsA induced VEGF mRNA and protein expressions at 7 and 28 days in LSD rats. At 7 days, CsA up-regulated the expression of Flt-1 and KDR/Flk-1 receptors; however, at 28 days, Flt-1 remained unchanged whereas KDR/Flk-1 expression declined. In NSD rats, in which the lesion did not develop, the expression of VEGF and its receptors remained similar to control. CONCLUSIONS: What causes VEGF to be up-regulated remains unclear. Further studies are needed to study the role of hypoxia and other cytokines in relation to VEGF in this model.     

高孕激素状态下促排卵与拮抗剂方案应用于卵巢低反应患者的促排卵疗效比较

目的比较高孕激素状态下促排卵方案与拮抗剂方案在卵巢低反应(POR)患者中应用的临床效果。方法回顾分析2015年10月至2016年10月在我院生殖中心行IVF/ICSI助孕的236例卵巢低反应患者的临床资料,其中116例采用高孕激素状态促排卵(PPOS)方案,120例采用拮抗剂方案,比较两组患者的基本情况及促排卵结局。结果两组患者年龄、基础性激素水平等一般资料比较无显著差异(P0.05);两组患者Gn用量[(2 729.25±1 048.50)U vs.(2 769.75±1 055.25)U]、直径≥14mm卵泡数[(3.51±2.44)vs.(3.95±2.77)]、获卵数[(2.86±2.05)vs.(2.86±2.27)]、无可移植胚胎周期取消率(29.3%vs.30.0%)比较亦无统计学差异(P0.05)。PPOS方案组2PN受精率(60.8%vs.58.6%)、卵裂率(83.5%vs.82.3%)、优胚率(54.0%vs.46.7%)及取卵周期取消率(6.0%vs.5.8%)略高于拮抗剂组,但差异无统计学意义(P0.05),但PPOS方案组可移植胚胎率显著高于拮抗剂方案组(72.2%vs.60.9%,P0.05)。PPOS方案组与拮抗剂组的Gn天数[(11.11±4.09)vs.(9.73±2.68)]、早发LH峰发生率(0vs.6.7%)有统计学差异(P0.05)。结论高孕激素状态下促排卵方案在POR患者能较好地抑制早发LH峰,增加可移植胚胎率,对于POR患者是较为理想的促排卵方案。     

Integrative treatment in endometriosis

Objective: To study the effectiveness of Yu's Neiyi Recipe on endometriosis.Methods: Yu's Neiyi Recipe was administrated in 48 patients with endometriosis (including 20 infertile cases). The clinical manifestations, ultrasonography, basal body temperature (BBT) and hormonal levels were observed before and 3 months after the treatment. The pregnancy outcome was followed up within 2 years.Results: Efficacy of Yu's Neiyi Recipe in the 48 cases was 93.75% within 3 months of treatment, 15 of the 20 infertile women become pregnant (80%) within 2 years. After the treatment, scores of dysmenorrhea, chronic pelvic pain or dyspareunia, and the size of ovarian endometrioma obviously decreased (P<0.01, P<0.01, P<0.01), meanwhile, the BBT type and high phase score (HPS) significantly improved (P<0.05). During herbal treatment, serum high PRL levels significantly dropped (P<0.05), and serum LH and E2 levels elevated significantly (P<0.05).Conclusions: Yu's Neiyi Recipe not only relieves the symptoms and signs of patients with endometriosis significantly, but also shows a good regulation on patients' ovarian function to improve the rate of ovulation and pregnancy.     

Expression of Trem-1 in the patients with endometriosis

Objective:Trem(triggering receptor expressed on myeloid cells)-1 is a novel activating receptor of the Ig super-family.In this study,we evaluated the levels of soluble Trem-1(sTrem-1)in peritoneal fluid(PF)and serum,to in-vestigate the role of Trem-1 in the pathogenesis and pathophysiology of endometriosis(EMS).Methods:The contents of sTrata-1 protein in sara and peritoneal fluid were measured in 21 women with EMS and 11 women without EMS by ELISA,and the expression of Trem-1 mRNA in the peripheral leucocytes in 6 women with EMS and 6 women without EMS by real-time PCR as well as RT-PCR.The expression of Trem-1 pro-tein in the ectopic endometrium of 6 women with EMS and without EMS was evaluated by immunohistochemical a-nalysis.Results:The concentration of strata-1 protein in PF from women with and without EMS were 33.60± 5.14 ng/ml and 14.60±3.13 ng/ml,respectively.There were significant differences in strata-1 levels in PF and serum between EMS patients and control(P<0.01).The expression of Tram-1 mRNA in the women with and without EMS was 0.77 ± 0.02 and 0.70±0.04,respectively.There was also significant difference between the two groups(P<0.05).But,there was not positive expression of Trem-1 in the ectopic endometrium.Conclusion:The expression of Trem-1 in women with EMS was up-regulated,suggesting that Tram-1 might play some important role in the pathogenesis and pathophysiology of EMS.     

The Kinase Insert Domain-containing Receptor (KDR) is Regulated by Shear Stress

Objective : Intimal hyperplasia develops in areas with low shear stress. The aim of the present study was to investigate if the mRNA expression of vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase-1 receptor (Flt-1) and kinase insert domain-containing receptor (KDR) is regulated by shear stress. Design : Endothelial cells from human umbilical veins were in an in vitro system subjected to different levels of shear stress during 1 and 12 h. The mRNA expression of VEGF, Flt-1 and KDR was measured with RT-PCR. eNOS served as positive control and actin as housekeeping gene. Results : The KDR expression was isolated upregulated 3-4 times after 12 h exposure to high shear stress. Conclusion : The genetic expression of KDR is upregulated by shear stress and this response is time dependent.     

The kinase insert domain-containing receptor (KDR) is regulated by shear stress

OBJECTIVE: Intimal hyperplasia develops in areas with low shear stress. The aim of the present study was to investigate if the mRNA expression of vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase-1 receptor (Flt-1) and kinase insert domain-containing receptor (KDR) is regulated by shear stress. DESIGN: Endothelial cells from human umbilical veins were in an in vitro system subjected to different levels of shear stress during 1 and 12 h. The mRNA expression of VEGF, Flt-1 and KDR was measured with RT-PCR. eNOS served as positive control and actin as housekeeping gene. RESULTS: The KDR expression was isolated upregulated 3-4 times after 12 h exposure to high shear stress. CONCLUSION: The genetic expression of KDR is upregulated by shear stress and this response is time dependent.